J Vet Sci.  2012 Sep;13(3):287-292. 10.4142/jvs.2012.13.3.287.

Protective effects of recombinant Brucella abortus Omp28 against infection with a virulent strain of Brucella abortus 544 in mice

• Lim JJ
• Kim DH
• Lee JJ
• Kim DG
• Min W
• Lee HJ
• Rhee MH
• Kim S

• KMID: 1389768
• DOI: http://doi.org/10.4142/jvs.2012.13.3.287

Abstract

The outer membrane proteins (OMPs) of Brucella (B.) abortus have been extensively studied, but their immunogenicity and protective ability against B. abortus infection are still unclear. In the present study, B. abortus Omp28, a group 3 antigen, was amplified by PCR and cloned into a maltose fusion protein expression system. Recombinant Omp28 (rOmp28) was expressed in Escherichia coli and was then purified. Immunogenicity of rOmp28 was confirmed by Western blot analysis with Brucella-positive mouse serum. Furthermore, humoral- or cell-mediated immune responses measured by the production of IgG1 or IgG2a in rOmp28-immunized mice and the ability of rOmp28 immunization to protect against B. abortus infection were evaluated in a mouse model. In the immunogenicity analysis, the mean titers of IgG1 and IgG2a produced by rOmp28-immunized mice were 20-fold higher than those of PBS-treated mice throughout the entire experimental period. Furthermore, spleen proliferation and bacterial burden in the spleen of rOmp28-immunized mice were approximately 1.5-fold lower than those of PBS-treated mice when challenged with virulent B. abortus. These findings suggest that rOmp28 from B. abortus is a good candidate for manufacturing an effective subunit vaccine against B. abortus infection in animals.

Keyword

Brucella abortus; immunization; rOmp28; vaccine

MeSH Terms

Animals
Antibodies, Bacterial/blood
Blotting, Western/veterinary
Brucella Vaccine/*immunology
Brucella abortus/*immunology
Brucellosis, Bovine/*immunology/microbiology/*prevention & control
Cattle
Cloning, Molecular
Electrophoresis, Polyacrylamide Gel/veterinary
Enzyme-Linked Immunosorbent Assay/veterinary
Female
Immunization/veterinary
Immunoglobulin G/blood
Immunoglobulin Isotypes/blood
Membrane Proteins/genetics/*immunology
Mice
Mice, Inbred BALB C
Models, Animal
Recombinant Proteins/genetics/immunology
Vaccines, Subunit/immunology

Figure

• 

  Fig. 1 Expression of recombinant Brucella (B.) abortus Omp28 (rOmp28) via a maltose fusion protein (pMAL) expression system. Total bacterial cell lysate proteins were separated by 10% SDS-PAGE and stained with Coomassie Brilliant Blue (A). The proteins were then transferred to a membrane and incubated with Brucella-positive mouse serum (B). pMAL (arrowheads) and rOmp28 (arrows) are indicated. M: marker, Lane 1: pMAL, Lane 2: rOmp28.

• 

  Fig. 2 Kinetics of the immune response elicited by immunization with rOmp28. The IgG1 (A) and IgG2a (B) log titers were determined by ELISA. Significant differences between the rOmp28-immunized and control (PBS-treated) mice are indicated by asterisks (*p < 0.05 and ***p < 0.001).

• 

  Fig. 3 Spleen weight of mice treated with PBS or immunized with rOmp28 and infected intraperitoneally with B. abortus. Spleen weights of the mice 10 days post-infection are shown. Data are presented as the mean ± SD (n = 5 per group). Spleen weight differed significantly between the rOmp28-immunized group and control (PBS-treated) animals as indicated by asterisks (**p < 0.01).

• 

  Fig. 4 Bacterial proliferation in the spleen of mice treated with PBS or immunized with rOmp28 and infected intraperitoneally with B. abortus. The numbers of viable bacteria recovered from spleens of the mice 10 days post-infections are shown. Data are presented as the mean ± SD (n = 5 per group). Bacteria proliferation differed significantly between the rOmp28-immunized group and control (PBS-treated) animals as indicated by asterisks (**p < 0.01).

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